• The Korea Journal of Herbology
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• v.33 no.3
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• pp.25-35
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• 2018

Objectives : Lepidii seu Descurainiae Semen has been frequently adulterated with the seeds of several inauthentic plant species. However, the accurate identification of these plant seeds is very difficult. To develop a reliable genetic authentication tool for Lepidii seu Descurainiae Semen, we analyzed matK sequence. Methods : To obtain the matK sequences of plant materials, genomic DNA was extracted from 24 samples and PCR amplification was carried out using matK-AF/matK-8R universal primer set and matK-LDSF/matK-LDSR primer set. For identifying species-specific nucleotides and phylogenetic analysis, matK regions were sequenced and comparatively analyzed by the ClustalW and Maximum Likelihood method. Results : We developed a new primer set to amplify matK region in Lepidii seu Descurainiae Semen and closely related plant samples. From the comparative analysis of matK sequences, we identified species-specific marker nucleotides for D. sophia, L. apetalum, L. latifolium, E. cheiranthoides, E. macilentum, and D. nemorosa, respectively. Furthermore, phylogenetic analysis revealed clear classification depending on the species. These results indicated that the matK sequence obtained a new primer set in this study was useful to identify Lepidii seu Descurainiae Semen in species level. Conclusions : We developed a primer set and identified species-specific marker nucleotides enough to distinguish authentic Lepidii seu Descurainiae Semen and adulterants at the species level based on the matK sequences. These genetic tool will be useful to prevent adulteration and to standardize the quality of Lepidii seu Descurainiae Semen.

• The Korea Journal of Herbology
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• v.34 no.5
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• pp.13-20
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• 2019

Objectives : To ensure the safety, quality and pharmacological efficacy of Batryticatus Bombyx, it is important to discriminate with adulterants. In Korean Herbal Pharmacopoeias (KHP), the authentic species of Batryticatus Bombyx is defined only Bombyx mori. Therefore, the aim of this study is establishment of PCR assay method using the sequence characterized amplified region (SCAR) marker based on COI DNA barcode for discriminating six species related to Batryticatus Bombyx. Methods : Seventeen samples of six species (Bombyx mori, Bombyx mandarina, Rhodinia fugax, Oberthueria caeca, Actias artemis, and Caligula japponica) were collected from different habitate and nucleotide sequences of cytochrome c oxidase subunit I(COI) barcode regions were analyzed by Sanger sequencing methods. To develop SCAR-based PCR assay method, we designed species-specific primers based on COI sequence variabilities and verified those specificities using 17 samples of six species as well as commercial herbal medicines. Results : In comparative multiple analysis of COI sequences, six species were distinguished by species-specific nucleotides at the species level. To develop rapid and reliable PCR assay method for genetic authentication of Batryticatus Bombyx, therefore, we designed species-specific SCAR primers based on these nucleotide sequences and confirmed those specificities. Using these SCAR primers, We also established simple conventional PCR assay method using these SCAR primers at the species level. Conclusions : The comparative analysis of COI sequences and SCAR-based PCR assay methods represented equal results for distinguishing authentic Batryticatus Bombyx and adulterations at the species level. Therefore, our results are expected protecting adulteration of herbal medicine Batryticatus Bombyx.

• Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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• 2001.06a
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• pp.1246-1246
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• 2001

The quality of meat is highly variable in many properties. This variability originates from both animal production and meat processing. At the pre-slaughter stage, animal factors such as breed, sex, age contribute to this variability. Environmental factors include feeding, rearing, transport and conditions just before slaughter (Hildrum et al., 1995). Meat can be presented in a variety of forms, each offering different opportunities for adulteration and contamination. This has imposed great pressure on the food manufacturing industry to guarantee the safety of meat. Tissue and muscle speciation of flesh foods, as well as speciation of animal derived by-products fed to all classes of domestic animals, are now perhaps the most important uncertainty which the food industry must resolve to allay consumer concern. Recently, there is a demand for rapid and low cost methods of direct quality measurements in both food and food ingredients (including high performance liquid chromatography (HPLC), thin layer chromatography (TLC), enzymatic and inmunological tests (e.g. ELISA test) and physical tests) to establish their authenticity and hence guarantee the quality of products manufactured for consumers (Holland et al., 1998). The use of Near Infrared Reflectance Spectroscopy (NIRS) for the rapid, precise and non-destructive analysis of a wide range of organic materials has been comprehensively documented (Osborne et at., 1993). Most of the established methods have involved the development of NIRS calibrations for the quantitative prediction of composition in meat (Ben-Gera and Norris, 1968; Lanza, 1983; Clark and Short, 1994). This was a rational strategy to pursue during the initial stages of its application, given the type of equipment available, the state of development of the emerging discipline of chemometrics and the overwhelming commercial interest in solving such problems (Downey, 1994). One of the advantages of NIRS technology is not only to assess chemical structures through the analysis of the molecular bonds in the near infrared spectrum, but also to build an optical model characteristic of the sample which behaves like the “finger print” of the sample. This opens the possibility of using spectra to determine complex attributes of organic structures, which are related to molecular chromophores, organoleptic scores and sensory characteristics (Hildrum et al., 1994, 1995; Park et al., 1998). In addition, the application of statistical packages like principal component or discriminant analysis provides the possibility to understand the optical properties of the sample and make a classification without the chemical information. The objectives of this present work were: (1) to examine two methods of sample presentation to the instrument (intact and minced) and (2) to explore the use of principal component analysis (PCA) and Soft Independent Modelling of class Analogy (SIMCA) to classify muscles by quality attributes. Seventy-eight (n: 78) beef muscles (m. longissimus dorsi) from Hereford breed of cattle were used. The samples were scanned in a NIRS monochromator instrument (NIR Systems 6500, Silver Spring, MD, USA) in reflectance mode (log 1/R). Both intact and minced presentation to the instrument were explored. Qualitative analysis of optical information through PCA and SIMCA analysis showed differences in muscles resulting from two different feeding systems.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.12
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• pp.1867-1872
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• 2010

Microbe and quality changes of vacuum-packaged ready-to-eat lettuce were analyzed. While the vacuumpackaged lettuce after chlorine sanitizer were stored at $5^{\circ}C$, $15^{\circ}C$, and $25^{\circ}C$ for 7 days, viable numbers of total aerobic bacteria (TAB), coliform, E. coli, food-borne pathogens and lactic acids bacteria (LAB) were counted with gas production and sensory evaluation. Before the storage, only TAB of 2 log CFU/g and coliform of 1 log CFU/g were detected and LAB was not detected. TAB, coliform and LAB increased by 1 log CFU/g at $5^{\circ}C$ for 7 days without any detection of the pathogens. Sensory evaluations for off-flavour and crispness dropped to half the best value at 5 day storage. TAB and coliform increased by 3 log CFU/g and 2 log CFU/g, respectively, but LAB grew very actively by 4 log CFU/g under anaerobic environment and only B. cereus were detected after enrichment of the lettuce at $15^{\circ}C$ for 3 days. The evaluations for off-flavour and crispness were half the best value for 3 days. However, TAB and coliform increased by 3 log CFU/g, 1 log CFU/g, and 4 log CFU/g, respectively only at 1 day storage under $25^{\circ}C$. Also B. cereus were detected after enrichment and the sensory evaluation were half the best value within 1 day storage. Therefore, preservation at the lowest temperature possible is required for growth inhibition of the bacteria contaminated in the lettuce. Interestingly, LAB among them grew most actively under the anaerobic condition and the adulteration of lettuce might be closely related with the growth of LAB.

• Journal of Food Hygiene and Safety
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• v.31 no.1
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• pp.28-35
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• 2016

Species identification of animal tissues in meat products is an important issue to protect the consumer from illegal and/or undesirable adulteration; for economic, religious and health reasons. In this reason, accurate analytical methods are needed for the labeling of meat products with requiring simple and fast procedure. Recently, applications of PCR in food analysis have been increased because of their simplicity, specificity and sensitivity. Therefore, in this study, a multiplex PCR assay was developed for the simultaneous identification of eight species of cow, pig, chicken, duck, goat, sheep, horse and turkey from raw meats. The primers were designed in different regions of mitochondrial 16S RNA after alignment of the available sequences in the GenBank database. Two multiplex primer sets were designed as Set 1 (cow, pig, chicken, duck) and Set 2 (goat, sheep, horse, turkey), respectively. Total 274 samples from cow (n = 55), pig (n = 30), chicken (n=30), and duck (n = 30), goat (n = 40), sheep (n = 33), horse (n = 41), and turkey (n = 15) were tested. The primers generated specific fragments of 94, 192, 279, 477 bp (pig, chicken, cow, duck), 670, 271, 152, 469 bp (goat, sheep, horse, turkey) lengths for eight species, respectively. The animal species specificity was 100% in all eight samples in the multiplex PCR assay. The detection limit of the multiplex PCR assay showed from 100 fg to 1 pg of template DNA from extracted from raw meats. When applying multiplex PCR assays to sample from pork/beef and pork/chicken, beef/chicken tested raw mixed meats and heat-treated ($83^{\circ}C$ for 30min, $100^{\circ}C$ for 20min, and $121^{\circ}C$ for 10min) mixtures, detection limit was 0.1% level beef, pork and pork in beef and chicken in pork and 1.0% level pork in chicken. This study suggest that the developed multiplex PCR assay can be used for rapid and simultaneous species identification of cow, pig, chicken, duck, goat, sheep, horse and turkey from meats.

• Journal of Food Hygiene and Safety
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• v.27 no.1
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• pp.68-73
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• 2012

In this study, a method was developed using molecular biological technique to distinguish an authenticity of meats for processed meat products. The genes for distinction of species about meats targeted at 12S or 16S genes in mitochondrial DNA and the species-specific primers were designed by that PCR products' size was around 200bp for applying to processed products. The target materials were 10 species of livestock products and it checked whether expected PCR products were created or not by electrophoresis after PCR using species-specific primers. The results of PCR for beef, pork, goat meat, mutton, venison, and horse meat were 131, 138, 168, 144, 191, and 142 bp each. The expected PCR products were confirmed at 281, 186, 174, and 238 bp for chicken, duck, turkeymeat, and ostrich. Also, non-specific PCR products were not detected in similar species by species-specific primers. The method using primers developed in this study confirm to be applicable for composite seasoning including beefs and processed meat products including pork and chicken. Therefore, this method may apply to distinguish an authenticity of meats for various processed products.

• Journal of Society of Preventive Korean Medicine
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• v.6 no.2
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• pp.66-85
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• 2002

Traditional herbal medicine is used extensively among the Korean populations, and other Asian countries employ similar therapies as well In recent years, extensive focus was laid on adulteration of the herbal medicine with heavy metals. This may be mainly due to a soil contamination by an environmental pollution. The objective of this study is to identify the contents of various heavy metals in the blood from OhJeokSan-Decoction (OD) treated-rats. For this study, 13 kinds of metals including essential and heavy metals, i.e. Al, As, Cd, Co, Cr, Cu, Fe, Hg, Mn, Ni, Pb, Se and Zn were analyzed by a slight modification of EPA methods and the following results are obtained. ; 1. There is no significant difference between the OD-treated groups and control group in liver, kidney, bone, brain, weight, food intake. 2. The amount of each metal analyzed in the liver are as follows; Al ; $0.235{\sim}4.215mg／kg$, As ; $0.103{\sim}0.461mg／kg$, Cd ; $0.005{\sim}0.010mg／kg$, Co ; $0.017{\sim}0.046mg／kg$, Cr ; $0.137{\sim}0.403mg／kg$, Cu ; $1.736{\sim}4.827mg／kg$, Fe ; $54.472{\sim}381.447mg／kg$, Hg ; not detected, Mn ; $1.159{\sim}2.803mg／kg$, Ni ; $0.007{\sim}0.095mg／kg$, Pb ; not detected, Se ; $0.682{\sim}1.887mg／kg$, Zn ; $10.213{\sim}26.119mg／kg$, by groups, respectively. In control and other experimental group, several metal (Co, Cu, Mn, Zn, As, Cr) has a significant difference, but in experimental I and other experimental II, III, IV, V groups, there are no significant difference. 3. The amount of each metal analyzed in the kidney are as follows; Al ; $1.712{\sim}31.230mg／kg$, As ; $0.062{\sim}0.439mg／kg$, Cd ; $0.010{\sim}0.062mg／kg$, Co ; $0.000{\sim}0.101mg／kg$, Cr ; $0.125{\sim}0.636mg／kg$, Cu ; $3.385{\sim}12.502mg／kg$, Fe ; $41.148{\sim}99.709mg／kg$, Hg ; $0.000{\sim}0.270mg／kg$, Mn ; $0.433{\sim}2.301mg／kg$, Ni ; $0.000{\sim}0.221mg／kg$, Pb ; $0.000{\sim}0.584mg／kg$, Se ; $0.540{\sim}1.600mg／kg$, Zn ; $8.775{\sim}17.060mg／kg$, by groups, respectively. The concentration of Cu, Se, Cr, and Hg are variated significantly in control and other experimental group, and Cu, Se, Cd, Cr are variated significantly in experimental I and other experimental II, III, IV, V groups. 4. The amount of each metal analyzed in the bone(tibia and fibula) are as follows; Al ; $9.557{\sim}119.464mg／kg$, As ; $0.139{\sim}12.250mg／kg$, Cd ; $0.000{\sim}0.295mg／kg$, Co ; $0.022{\sim}0.243mg／kg$, Cr ; $0.239{\sim}1.999mg／kg$, Cu ; $0.000{\sim}2.291mg／kg$, Fe ; $240.249{\sim}841.956mg／kg$, Hg ; $0.000{\sim}0.983mg／kg$, Mn ; $0.214{\sim}7.353mg／kg$, Ni ; $5.473{\sim}11.453mg／kg$, Pb ; $0.000{\sim}8.502mg／kg$, Se ; $0.000{\sim}3.005mg／kg$, Zn ; $61.158{\sim}195.038mg／kg$, by groups, respectively. The concentration of Se, Cd are variated significantly in control and other experimental groups, and Se is variated significantly in experimental I and other experimental II, III, IV, V groups. 5. Exceptionally several metal concentration is increased or decreased. but there is no significant harmful difference of metal concentration in the liver, kidney and bone, from the OD-treated-rats compared to those of the control group even if higher dosage($1{\sim}8$ times dosage of person) of OD was administered. Thus, it is expected that the herbal decoction in the traditional herbal medicine would not lay any burden on the body and the heavy metal toxins would not affect our physiological system. However, other kinds of herbal treatment, such as i.v. and i.p. should be considered in terms of metal toxicity in the body since the level of certain metal.